Interaction of Adriamycin and N-Trifluoroacetyladriamycin-14-Valerate with Cardiolipin-Containing Lipid Bilayers

Abstract

The interaction of adriamycin (ADR) and N-Trifluoroacetyladriamycin-14-valerate (AD 32) with cardiolipin (CL)-containing multilamellar vesicles was studied by high-sensitivity differential scanning calorimetry, using liposomes formed from either dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosphatidylglycerol (DPPG) containing small amounts of CL. the drugs partitioned into cardiolipin-containing neutral and acidic bilayers in a manner similar to that observed earlier with CL-free bilayers. the partition coefficient of adriamycin in bilayers of vesicles prepared from DPPG or DPPG in admixture with CL was much higher than that obtained with neutral DPPC vesicles or the DPPC together with CL. Under all conditions, AD 32 was essentially completely partitioned into the lipid phase of the investigated phospholipid membranes. As expected, addition of adriamycin to CL-containing vesicles did not significantly change the thermotropic behavior of these bilayers, whereas the fluidizing effect of AD 32 was directly related to the CL content of the vesicles. the multipeak transitions produced by the anthracyclines in pure DPPG bilayers were preserved in the presence of CL, but the endotherms were broader and slightly shifted to lower temperatures, a finding indicative of stronger interactions. Chlorpromazine (CPZ), which was used as a reference compound to compare the effects produced by the anthracyclines, was found to behave similarly to AD 32 and to be more effective than quinidine (QND), in agreement with their behavior in CL-free liposomes.     

Membrane fusion between liposomes composed of acidic phospholipids and neutral phospholipids induced by melittin: a differential scanning calorimetric study

Melittin-induced membrane fusion between neutral and acidic phospholipids was examined in liposome systems with a high-sensitivity differential scanning calorimeter. Membrane fusion could be detected by calorimetric measurement by observing thermograms of mixed liposomal lipids. The roles of hydrophobic and electrostatic interactions were investigated in membrane fusion induced by melittin. Melittin, a bee venom peptide, is composed of a hydrophobic region including hydrophobic amino acids and a positively charged region including basic amino acids. When phosphatidylcholine liposomes were prepared in the presence of melittin, reductions in the phase transition enthalpies were observed in the following order; dimyristoylphosphatidylcholine (DMPC) > dipalmitoylphosphatidylcholine (DPPC) > distearoylphosphatidylcholine (DSPC) > dielaidoylphosphatidylcholine (DEPC). The plase transition enthalpy of an acidic phospholipid, dipalmitoylphosphatidylserine (DPPS), was raised by melittin at low concentrations, then reduced at higher concentrations. DPPC liposomes prepared in melittin solution were fused with DPPS liposomes when the liposomal dispersions were mixed and incubated. Similar fusion was observed between dipalmitoylphosphatidylcholine and dimyristoylphosphatidic acid (DMPA) liposomes. These results indicate that a peptide including hydrophobic and basic regions can mediate membrane fusion between neutral and acidic liposomes by hydrophobic and electrostatic interactions.     

The characteristics of the radiation-initiated peroxidation of the phosphatidylcholine making up liposomes containing phospholipids which are susceptible to free-radical fragmentation

The regularities of accumulation of conjugated dienes and thiobarbituric acid (TBA)-reactive substances under gamma-irradiation of liposomes from rat liver phosphatidylcholine (PC) and its mixtures with the resistant to lipid peroxidation saturated phospholipids and bovine brain sphingomyelin (SM) were studied. It was established that the incorporation of negatively charged dipalmitoylphosphatidylglycerol (DPPG) and dipalmitoylphosphatidylethanol (DPPET) into lipid bilayer resulted in the increase of primary and secondary products of LPO, whereas neutral dipalmitoylphosphatidylcholine (DPPC) and SM involving in the phospholipid mixtures inhibited the peroxidation of PC. For anionic phospholipids, DPPG had more profound activating action on LPO, amongst the neutral phospholipids SM was more potent inhibitor of the reaction. Unlike DPPET and DPPC, DPPG and SM were subjected to free radical fragmentation on gamma-radiation. It is suggested that the intermediates and products of free radical fragmentation may modulate the progress of LPO.     

Effects of local anesthetics on membrane properties. I. Changes in the fluidity of phospholipid bilayers.

The effect of the local anesthetic dibucaine on the solid to liquid-crystalline phase transition in phospholipid vesicles was studied by calorimetry and fluorescence polarization. The partition coefficient (greater than 3000) of dibucaine in the membranes of vesicles prepared from acidic phospholipids was more than 20 times higher than in neutral phospholipid membranes under the same conditions. Calorimetric measurements on vesicles prepared form acidic phospholipids (bovine brain phosphatidylserine; dipalmitoylphosphatidylglycerol) showed that dibucaine (1 with 10(-4) M) produced a significant reduction in the gel-liquid crystalline transition temperature (Tc). This fluidizing effect of dibucaine on acidic phospholipid membranes was even more marked in the presence of Ca2+. In contrast, dibucaine at the same concentration did not alter the Tc of neutral phospholipids (dipalmitoylphosphatidylcholine). Significant increase in the fluidity of neutral phospholipid membranes occurred only at higher dibucaine concentrations (2 with 10(-3) M). Measurements of the fluorescence polarization and lifetime of the probe, 1,6-diphenylhexatriene, in acidic phospholipid vesicles revealed that dibucaine (1 with 10(-4) M) caused an increase in the probe rotation rate indicating an increase in the fluidity of the phospholipid membranes. A good correlation was obtained between fluorescence polarization data on dibucaine-induced changes in membrane fluidity and calorimetric measurements on vesicles of the same type.     

Quinidine is a strong perturber of acidic phospholipid bilayer order and fluidity

The effect of an antiarrhythmic drug, quinidine, on the organization of model phospholipid membranes was studied by the spin-labeling technique. Quinidine strongly perturbs the molecular organization of lipid bilayers prepared from acidic phospholipids (phosphatidylserine, phosphatidic acid) and has only a slight effect on neutral phosphatidylcholine membranes. The interaction of the drug with acidic phospholipids manifests itself in a pronounced increase in the order parameter of the region close to the polar surface of the bilayer and in some decrease in its inner hydrocarbon core fluidity. It is suggested that the perturbation in the organization of membrane lipids may contribute to the mechanisms by which quinidine exerts its pharmacological effects.     

Effects of 2.45-GHz microwave and 100-MHz radiofrequency radiation on liposome permeability at the phase transition temperature

Large unilamellar dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) liposomes loaded with an aqueous chemotherapeutic drug, cytosine arabinofuranoside (ARA-C), were exposed for 30 min to 60 W/kg continuous-wave (CW) 100-MHz or 2.45-GHz radiation in vitro at temperatures between 37 degrees C and 43 degrees C. Liposomes were exposed in HEPES buffer or in HEPES buffer supplemented with 44% by volume fetal calf serum (FCS). Characteristic phase transition responses were detected in the range of 39 degrees C to 40 degrees C with the presence of FCS, increasing maximum % release of 3H-ARA-C by 20% relative to HEPES suspension. Neither frequency of electromagnetic radiation had any detectable effect on liposome permeability or the location of the phase transition in the presence or absence of FCS.     

Interaction of wasp venom mastoparan with biomembranes

Mastoparan-induced changes in the K+ permeability of rat peritoneal mast cells, human erythrocytes, Staphylococcus aureus and Escherichia coli were examined. Mastoparan did not efficiently increase the K+ permeability of cells except for S. aureus. The release of membrane phospholipids was also observed from S. aureus cells in the concentration range of the permeability enhancement. Mastoparan stimulated histamine release from mast cells, independently of a small efflux of K+. Mastoparan became markedly effective to E. coli cells whose outer membrane structure was chemically disrupted beforehand, showing that the peptide can enhance the permeability of the cytoplasmic membranes of both Gram-positive and -negative bacteria. In experiments using liposomes, mastoparan increased the permeability of the liposomes composed of egg phosphatidylethanolamine and egg phosphatidylglycerol, which are the lipid constituents of the cytoplasmic membrane of E. coli cells, while it showed a weak activity to the liposomes composed of egg phosphatidylcholine and cholesterol. The latter result related closely to the fact that this peptide acted weakly on erythrocytes and mast cells in which acidic lipids constitute a minor portion. Mastoparan decreased the phase transition temperature of dipalmitoylphosphatidylglycerol liposomes, but it did not affect that of dipalmitoylphosphatidylcholine liposomes. These results indicate that mastoparan penetrated into membranes mainly containing acidic phospholipids and disrupted the membrane structure to increase the permeability. The action of the wasp venom mastoparan was compared with that of a bee venom melittin.     

Surface areas of lipid membranes

Upon photolysis, alkyl pentacyanocobaltate complexes generate alkyl radicals which react rapidly and specifically with nitroxide radicals, and which do not penetrate phospholipid bilayers. By measuring the loss of paramagnetic resonance signal intensity when multilamellar liposomes containing a small amount of spin-labeled lipid are exposed to these radicals, we have measured the proportion of lipid on the external surface of liposomes. We have shown that liposomes prepared under specified conditions from dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, and binary mixtures of dipalmitoylphosphatidylcholine and cholesterol all have the same proportion of external lipid.     

Temperature-dependent permeability of large unilamellar liposomes

The temperature-dependent drug leakage from liposomes composed of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol (4:1, by weight) was studied. Experiments were performed in Hepes buffer and 50% fetal calf serum. Large unilamellar liposomes were formed by the reverse phase evaporation process and extruded through a series of polycarbonate membranes with pore sizes of 0.4, 0.2, 0.1 and 0.08 μm. The release of the water soluble radioisotopes cytosine 1-β-D-[³H]arabinofuranoside and [³H]inulin from the aqueous compartment of these liposomes was measured as a function of time and temperature. Both radioisotopes were released at temperatures near 42°C, the solid-to-liquid-crystalline phase transition temperature of these lipids. The percent drug release decreased as the size of the liposomes was reduced. This effect was more pronounced in Hepes buffer than serum. The release of both radioisotopes was greatest at 40°C in Hepes buffer and at 43°C in 50% fetal calf serum. In addition, the rate of drug release was much faster in serum than in buffer. These results suggest that different drug release processes are occurring in buffer and in serum.     

Changes of transition temperatures of phosphatidylcholine and phosphatidylglycerol in presence of amphiphilic drugs

The transition temperature of phosphatidylglycerol (PG) was reduced to lower temperatures in presence of propranolol, imipramine, amitriptyline and chlorpromazine. This effect was dependent on drug concentration and was smallest with propranolol. The fluidizing effect, however, increased from propranolol to chlorpromazine according to the octanol/H2O partition coefficients. When the two phospholipids PG and phosphatidylcholine (PC) were compared, the presence of drug lead to a more pronounced reduction of the transition temperature in the case of the acidic phospholipid than in the case of the neutral one.     

Reconstitution of sodium channels in large liposomes formed by the addition of acidic phospholipids and freeze-thaw sonication

Phosphatidylcholine (PC) alone or with phosphatidylethanolamine (PE) are sufficient for the reconstitution of Na+ channels in planar lipid bilayers. However, when Na+ channels were first reconstituted into liposomes using the freeze-thaw-sonication method, addition of acidic phospholipids, such as phosphatidylserine (PS), to the neutral phospholipids was necessary to obtain a significant toxin-modulated 22Na uptake. To further investigate the acidic phospholipid effect on reconstitution into liposomes, Na+ channels purified from Electrophorus electricus electrocytes were reconstituted into liposomes of different composition by freeze-thaw sonication and the effect of batrachotoxin and tetrodotoxin on the 22Na flux was measured. The results revealed that, under our experimental conditions, the presence of an acidic phospholipid was also necessary to obtain a significant neurotoxin-modulated 22Na influx. Though neurotoxin-modulated 22Na fluxes have been reported in proteoliposomes made with purified Na+ channels and PC alone, the 22Na fluxes were smaller than those found using lipid mixtures containing acidic phospholipids. Electron microscopy of negatively stained proteoliposomes prepared with PC, PC/PS (1:1 molar ratio), and PS revealed that the acidic phospholipid increases the size of the reconstituted proteoliposomes. The increment in size caused by the acidic phospholipid, due to the associated increase in internal volume for 22Na uptake and in area for Na+ channel incorporation, appears to be responsible for the large neurotoxin-modulated 22Na fluxes observed.     

Binding of adriamycin to liposomes as a probe for membrane lateral organization.

A stopped-flow spectrofluorometer equipped with a rapid scanning emission monochromator was utilized to monitor the binding of adriamycin to phospholipid liposomes. The latter process is evident as a decrease in fluorescence emission from a trace amount of a pyrene-labeled phospholipid analog (PPDPG, 1-palmitoyl-2-[(6-pyren-1-yl)]decanoyl-sn-glycero-3-phospho-rac-++ +glyce rol) used as a donor for resonance energy transfer to adriamycin. For zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes, fluorescence decay was slow, with a half-time t1/2 of approximately 2 s. When the mole fraction of the acidic phospholipid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-glycerol (POPG), was increased to XPG >/= 0.04, the decay of fluorescence became double exponential, and an additional, significantly faster process with t1/2 in the range between 2 and 4 ms was observed. Subsequently, as XPG was increased further, the amplitude of the fast process increased, whereas the slower process was attenuated, its t1/2 increasing to 20 s. Increasing [NaCl] above 50 mM or [CaCl2] above 150 microM abolished the fast component, thus confirming this interaction to be electrostatic. The critical dependence of the fast component on XPG allows the use of this process to probe the organization of acidic phospholipids in liposomes. This was demonstrated with 1, 2-palmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes incorporating PPDPG (XPPDPG = 0.03), i.e., conditions where XPG in fluid bilayers is below the required threshold yielding the fast component. In keeping with the presence of clusters of PPDPG, the fast component was observed for gel-state liposomes. At approximately 34 degreesC (i.e., 6 degrees below Tm), the slower fluorescence decay also appeared, and it was seen throughout the main phase transition region as well as in the liquid-crystalline state. The fluorescence decay behavior at temperatures below, above, and at the main phase transition temperature is interpreted in terms of thermal density fluctuations and an intermediate state between gel and liquid-crystalline states being involved in the phospholipid main phase transition. This is the first observation of a cluster constituted by acidic phospholipids controlling the membrane association of a drug.     

Antioxidant activity of probucol and its effects on phase transitions in phosphatidylcholine liposomes

The effect of probucol on the phase behavior of dimyristoylphosphatidylcholine (DMPC) was examined by fluorescence polarization and differential scanning calorimetry (DSC). Probucol broadens and shifts the temperature of the main phase transition of DMPC liposomes as measured by fluorescence polarization with diphenylhexatriene and trimethyl-ammonium-diphenylhexatrine at concentrations as low as 5 mole%. As measured by DSC, probucol reduces the transition temperature of the gel----liquid-crystalline phase transition of DMPC by approx. 2 C degrees at all concentrations above about 5 mole% probucol and eliminates the pretransition at less than 1 mole%. In addition, the phase transition of DMPC is broadened and the enthalpy of the transition reduced by approx. 50%. Even at high concentrations of probucol, the gel----liquid-crystalline phase transition of DMPC is not eliminated. Similar effects are observed with dipalmitoylphosphatidylcholine liposomes. Based on these DSC measurements, measurements of the melting of probucol in dry mixtures with DMPC and observations of probucol mixtures with DMPC under polarizing optics, the maximum solubility of probucol in DMPC is approx. 10 mole%. This concentration exceeds that required (approx. 0.5 mole%) to prevent peroxidation of 10 mole% arachidonic acid in DMPC liposomes for 30 min in the presence of 0.05 mM Fe(NH4)(SO4)2 at 4 degrees C. Thus, probucol has a limited solubility in saturated phosphatidylcholine bilayers, but is an effective antioxidant at concentrations lower than its maximum solubility.     

Effects of local anesthetics on membrane properties I. Changes in the fluidity of phospholipid bilayers

The effect of the local anesthetic dibucaine on the solid to liquid-crystalline phase transition in phospholipid vesicles was studied by calorimetry and fluorescence polarization. The partition coefficient (> 3000) of dibucaine in the membranes of vesicles prepared from acidic phospholipids was more than 20 times higher than in neutral phospholipid membranes under the same conditions. Calorimetric measurements on vesicles prepared form acidic phospholipids (bovine brain phosphatidylserine; dipalmitoylphosphatidylglycerol) showed that dibucaine (1 · 10⁻⁴M) produced a significant reduction in the gel-liquid crystalline transition temperature (T_c). This fluidizing effect of dibucaine on acidic phospholipid membranes was even more marked in the presence of Ca²⁺. In contrast, dibucaine at the same concentration did not alter the T_c of neutral phospholipids (dipalmitoylphosphatidylcholine). Significant increase in the fluidity of neutral phospholipid membranes occurred only at higher dibucaine concentrations (2 · 10⁻³M. Measurements of the fluorescence polarization and lifetime of the probe, 1,6-diphenylhexatriene, in acidic phospholipid vesicles revealed that dibucaine (1 · 10⁻⁴M caused an increase in the probe rotation rate indicating an increase in the fluidity of the phospholipid membranes. A good correlation was obtained between fluorescence polarization data on dibucaine-induced changes in membrane fluidity and calorimetric measurements on vesicles of the same type.     

Interaction of the cytoskeletal component vinculin with bilayer structures analyzed with a photoactivatable phospholipid

The cytoskeletal component vinculin has been proposed to act as an actin-plasma membrane linker. In order to demonstrate a possible direct interaction of vinculin with bilayers, photolabeling with a phospholipid generating a highly reactive carbene was used. This phosphatidylcholine analogue (1-palmitoyl-2-[10-[4-[(trifluoromethyl)diazirinyl]phenyl]-[3H] 9-oxaundecanoyl]-sn-glycero-3-phosphocholine), with the photoactivatable diazirine group on its apolar portion, has been shown to label selectively membrane-embedded domains of membrane proteins. Vinculin is significantly labeled upon incubation and photolysis with liposomes containing trace amounts of this photoactivatable phospholipid, but only when the liposomes also contain acidic phospholipids. Labeling of vinculin is markedly increased (5-17-fold) by all acidic phospholipids tested so far (30%, w/w), compared to labeling in neutral phospholipids. Labeling is high at low ionic strength, but significant vinculin labeling can still be observed at physiological salt concentrations and acidic phospholipid content of the membrane. Our results provide evidence that vinculin inserts into the hydrophobic part of the bilayer by interacting with acidic phospholipids. A similar interaction may be of importance in vivo.     

Physical properties of phosphatidylcholine-phosphatidylinositol liposomes in relation to a calcium effect

Physical properties of binary mixtures of dipalmitoylphosphatidylcholine and yeast phosphatidylinositol were studied by ESR analysis using TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl) and lipid spin probes, freeze-fracture electronmicroscopy and particle microelectrophoresis, and they were compared with those of phosphatidylcholine/bovine brain phosphatidylserine mixtures. The phase diagram of the binary mixtures of dipalmitoylphosphatidylcholine and phosphatidylinositol was obtained from the thermal features of TEMPO spectral parameter in the lipid mixtures. The phase diagram provided evidence that these two phospholipids in various combinations were miscible in the crystalline state. The addition of 10 mM Ca2+ slightly shifted the phase diagram upward. TEMPO titration of the binary mixture of dipalmitoylphosphatidylcholine and bovine brain phosphatidylserine revealed that 10 mM Ca2+ caused the complete phase separation of this lipid mixture. Studies of phase separations using phosphatidylcholine spin probe manifested that 10 mM Ca2+ induced almost complete phase separation in egg yolk phosphatidylcholine/bovine brain phosphatidylserine mixtures but only slight phase separation in egg yolk phosphatidylcholine/yeast phosphatidylinositol mixtures. However, some phase changes around the fluidus and the solidus curves were visualized by the freeze-fracture electronmicroscopy. The molecular motion of lipid spin probe was decreased by the addition of Ca2+ in the liposomes containing phosphatidylinositol. The temperature dependence of electrophoretic mobility was also examined in the absence and presence of 1 mM Ca2+. Liposomes of dipalmitoylphosphatidylcholine-phosphatidylinositol (90 : 10, mol/mol) exhibited a clear transition in the thermal features of electrophoretic mobilities. Raising the phosphatidylinositol content up to 25 mol% rendered the transition broad and unclear. The addition of 1 mM Ca2+ decreased the electrophoretic mobility but did not change its general profile of the thermal dependence. These results suggest that the addition of calcium ions induced a small phase change in the binary mixture of phosphatidylcholine and phosphatidylinositol while Ca2+ causes a remarkable phase separation in phosphatidylcholine/phosphatidylserine mixture. The physical role of phosphatidylinositol is discussed related to the formation of diacylglycerol.     

Liposomes, disks, and spherical micelles: aggregate structure in mixtures of gel phase phosphatidylcholines and poly(ethylene glycol)-phospholipids

Poly(ethylene glycol) (PEG) decorated lipid bilayers are widely used in biomembrane and pharmaceutical research. The success of PEG-lipid stabilized liposomes in drug delivery is one of the key factors for the interest in these polymer/lipid systems. From a more fundamental point of view, it is essential to understand the effect of the surface grafted polymers on the physical-chemical properties of the lipid bilayer. Herein we have used cryo-transmission electron microscopy and dynamic light scattering to characterize the aggregate structure and phase behavior of mixtures of PEG-lipids and distearoylphosphatidylcholine or dipalmitoylphosphatidylcholine. The PEG-lipids contain PEG of molecular weight 2000 or 5000. We show that the transition from a dispersed lamellar phase (liposomes) to a micellar phase consisting of small spherical micelles occurs via the formation of small discoidal micelles. The onset of disk formation already takes place at low PEG-lipid concentrations (<5 mol %) and the size of the disks decreases as more PEG-lipid is added to the lipid mixture. We show that the results from cryo-transmission electron microscopy correlate well with those obtained from dynamic light scattering and that the disks are well described by an ideal disk model. Increasing the temperature, from 25 degrees C to above the gel-to-liquid crystalline phase transition temperature for the respective lipid mixtures, has a relatively small effect on the aggregate structure.     

The physical state of membrane lipids modulates the activation of the first component of complement.

Activation of the first component of human complement (C1) by bilayer-embedded nitroxide spin label lipid haptens and specific rabbit antinitroxide antibody has been measured. The nitroxide spin label hapten was contained in host bilayers of either dimyristoyl phosphatidylcholine or dipalmitoyl phosphatidylcholine in the form of both liposomes and vesicles. At a temperature of 32 degrees C, which is intermediate between the hydrocarbon chain-melting temperatures of the two phospholipids, activation of C1 in such vesicles and liposomes is more efficient in the fluid membrane. Studies of C1 activation in binary mixtures of cholesterol and dipalmitoyl phosphatidylcholine indicate that the activation of C1 is not limited by the lateral diffusion of the lipid haptens in these membranes.     

Interaction of Adriamycin with Single and Multibilayer Dipalmitoylphosphatidylcholine Vesicles: Spin-Labeling and Calorimetric Study

Abstract

The effects of adriamycin on the organization of dipalmitoylphosphatidylcholine (DPPC) membranes alone or in the presence of 1 mol% cardiolipin (CL) in the form of single and multibilayer vesicles has been studied by spin-labeling, and by high sensitivity differential scanning calorimetry. With sonicated small unilamellar vesicles (SUVs), adriamycin increased the order parameter of 5-doxylstearate spin-labeled vesicles, an effect primarily observed in the gel phase of DPPC. thermal transition profiles, obtained by high-sensitivity differential scanning calorimetry and by 2,2,6,6-tetramethylpiperidinyl-l-oxy (TEMPO) partitioning studies, indicated that the drug induced aggregation and fusion of SUVs, especially at high drug-lipid molar ratios, and this phenomenon was further verified by negative-staining electron microscopy. the presence of J mol% CL in the lecithin bilayer markedly enhanced the effect of adriamycin on membrain order and fusion, especially under conditions of low ionic strength. the ordering effect of the drug was insensitive to the presence of other acidic phospholipids and was only partially inhibited by Ca²⁺ or Mg²⁺. In contrast to the small and highly-curved SUVs, however, the phase behavior of fused unilamellar vesicles (FUVs) or multilamellar vesicles (MLVs) was not significantly affected by the drug, suggesting that the bilayer curvature is an important factor in the interaction of the antibiotic with the corresponding bilayers. these findings demonstrate that changes in the bilayer packing configuration, due to differences in the radii of the curvature of the vesicles, must be considered in studies of adriamycin-lipid membrane interactions, as well as the phospholipid composition of the vesicles.     

Neutral Lipids Rigidify Unsaturated Acyl Chains in Senescing Membranes

Senescence in bean cotyledons is accompanied by a progressiveincrease in the proportion of gel phase lipid in cellular membranesthat can be attributed to qualitative changes in the neutrallipids. The resulting mixture of lipid phases leads to impairedmembrane function. Insight into the molecular basis for thisphenomenon has been gleaned from studies of the effects of theseneutral lipids on the phase properties of pure phospholipidmembranes. Induction of the gel phase, detectable as a risein the liquid-crystalline to gel phase transition temperature,was observed when neutral lipid from senescent membranes wasintroduced into liposomes of unsaturated phospholipids. Thetransition temperature for phosphatidylcholine rose from –6.5° C in control liposomes to 51 ° C when 25% (w/w) neutrallipid was present. A similar rise was obtained for dioleoylphosphatidylcholine.However, for dipalmitoylphosphatidylcholine, which is fullysaturated, the rise in transition temperature upon additionof neutral lipids was only 3 ° C. Thus the neutral lipidsin senescent membranes appear to selectively rigidify unsaturatedacyl chains. At least three types of compounds known to alterthe phase properties of lipid bilayers were detectable in theneutral lipid fraction, suggesting that rigidification reflectsthe concerted action of several neutral lipid components onunsaturated phospholipid. Key words: Senescence, phospholipid, lipid phase properties